Non-adhesive elastic gelatin matrices

ABSTRACT

The present invention is a substantially non-adhesive elastic gelatin matrix. The matrix is both non-adhesive to wounds, tissues and organs and is also elastic such that it is flexible. The matrix is a lyophilized mixture of protein(s), polymer(s), cross-linking agent(s) and optional plasticizer(s). The invention also provides methods for making the non-adhesive elastic gelatin matrix.

RELATED APPLICATIONS

This application is a continuation of U.S. patent application Ser. No.11/664,184, filed Aug. 8, 2008, now abandoned which corresponds to PCTInternational Application No. PCT/CA2005/000925, filed Jun. 15, 2005,which claims the benefit of U.S. Provisional Patent Application Ser. No.60/614,414, filed Sep. 30, 2004. The subject matter of theaforementioned applications is incorporated herein by reference in theirentireties.

FIELD OF THE INVENTION

The present invention relates to non-adhesive elastic matrices. Morespecifically, the present invention relates to non-adhesive cross-linkedelastic gelatin matrices, and the method of making the same. Thematrices can be provided in a variety of configurations not limited towound dressings, wound barriers, tissue and vascular coverings. Thematrices can also be made to incorporate a variety of pharmaceuticals,chemicals and other agents. Further, the non-adhesive elastic matricesof the present invention can be used alone, or in conjunction with othermaterials.

BACKGROUND TO THE INVENTION

Gelatin sheets, collagen sheets and sponges are a group of biomaterialsthat have been used extensively in the medical field. They are dry andabsorb and retain large amounts of water. This group of biomaterials isboth biocompatible and biodegradable causing little or no inflammation.These biomaterials are useful as wound dressings, artificial skinscaffolds and therapeutic drug delivery devices, whereby thebiomaterials can retain therapeutics and deliver such therapeutics toappropriate cells and tissues, as exemplified in Applicant's U.S. Pat.No. 6,475,516.

Gelatin and collagen sheets as provided by the prior art are typicallyinelastic and may have toxic properties. For example, U.S. Pat. No.3,491,760 describes a pliable, but inelastic, adhesive multilayer woundcovering composed of collagen or gelatin which is “foamed” using airbubbles, treated with a plasticizer (glycerol), cross-linked with a 4%glutaraldehyde solution, and the resulting gel film covered with anadhered cover layer. As is generally known collagen and gelatin matricescross-linked with glutaraldehyde show residual toxicity.

Gelatin sponge materials such as Gelfoam™, are also known. Thesematerials are absorbable, sterile and water insoluble and used tocontrol bleeding during surgery and may also be provided in a powderformat. The material is non-elastic. Another non-elastic gelatin spongecurrently used is Surgifoam™ Gelfilm™ is a non-elastic material obtainedfrom a formaldehyde cross-linked gelatin solution.

Hydrolyzed collagen, also known as gelatin, has been used as a vehiclefor the delivery of therapeutics when covalently attached to the surfaceof tubular medical devices such as catheters and stents as described inApplicant's U.S. Pat. Nos. 6,132,765 and 6,228,393. The material ishydrated and minimally elastic.

Several patents describe cross-linked inelastic collagen material suchas for example U.S. Pat. Nos. 4,703,108, 4,970,298, 5,550,187,5,744,545, and 6,132,765.

Sehal and Vijay (Anal. Biochem. 218: 87-91, 1994) describe a method forwater-soluble carbodiimide-mediated amidation by using1-ethyl-3-[3-dimethylaminopropyl carbodiimide (EDC)/N-hydroxysuccinimide (NHS). The method is known to cross-link collagen, gelatinand other proteins. However, the resultant matrices are not elastic.

Choi et al. (Biomaterials 20: 409-41, 1999) describe the fabrication ofa gelatin-containing artificial skin that contains gelatin and alginateand is cross-linked with EDC. A soluble sponge was immersed in asolution of acetone water (9:1 by volume) containing 20-100 mg EDC andcross-linked for 24 hr. The EDC when dissolved in water is stated to bedeactivated and rapidly loses its cross-linking ability and thus the EDCwas dissolved in 90% acetone. The matrix material produced is inherentlyinelastic.

In the pharmaceutical industry, soft elastic gelatin capsules or“Softgels” are made from a gelatin solution that is plasticized withpropylene glycol, sorbitol, glycerin or other approved mixtures.However, the gelatin in the softgel capsules is not cross-linked inorder that the capsules remain soluble when ingested. Furthermore,softgel capsules lack elasticity and resistance to the action ofdegradative processes such as increased temperature and mild enzymaticaction.

There is a need to produce gelatin matrices that are elastic, and haveappreciable tensile strength while not being prone to becoming hard andbrittle.

SUMMARY OF THE INVENTION

The present invention provides substantially non-adhesive,proteinaceous, elastic matrices for a variety of clinical uses. Morespecifically, the present invention provides substantially non-adhesive,elastic gelatin matrices for use in a variety of applications notlimited to, wound barriers, wound dressings, surgical dressings, wrapsand in therapeutic drug and/or chemical agent delivery. The presentinvention also provides a novel method for making the non-adhesive,elastic gelatin matrices of the invention.

In accordance with an aspect of the present invention is a non-adhesiveelastic gelatin matrix. The matrix is both non-adhesive and elastic suchthat it is flexible. The matrix is lyophilized such that it does notcontain substantial amounts of solvent and comprises a mixture ofcollagen or denatured collagen, biocompatible polymers, cross-linkingagent(s) and optional plasticizer.

In accordance with another aspect of the present invention is alyophilized non-adhesive elastic gelatin matrix.

In accordance with another aspect of the present invention is anon-adhesive elastic protein matrix, the matrix comprising;

-   -   a mixture of protein comprising collagen and/or gelatin,        biocompatible polymers, one or more cross-linking agents. In        aspects, the mixture may further comprise one or more        plasticizers. In other aspects, the mixture is lyophilized.

In aspects, collagen or denatured collagen, i.e. gelatin is used aloneas the protein component of the matrix. In alternative aspects, collagenand gelatin are used in combination as the protein component of thematrix.

In accordance with another aspect of the present invention is anon-adhesive elastic gelatin matrix, the matrix comprising;

-   -   a mixture of protein comprising collagen and/or gelatin and        biocompatible polymers comprising alginate, polyethylene glycol        and poly-L-lysine, wherein said polymers are cross-linked with        one or more cross-linking agents and said mixture is        lyophilized. In aspects, the mixture may further comprise one or        more plasticizers.

In aspects, the collagen can be used alone or denatured collagen, i.e.gelatin used alone or in combination.

In accordance with another aspect of the present invention, there isprovided a method for making a non-adhesive elastic gelatin matrix, themethod comprising:

-   -   lyophilizing a frozen solution comprising at least one protein,        at least one biocompatible polymer and at least one        cross-linking agent for a time effective to substantially remove        the solvent. In aspects, the solution also comprises a        plasticizer.

In accordance with still another aspect of the present invention is amethod for making a non-adhesive elastic gelatin matrix, the methodcomprising:

-   -   (a) heating a protein solution;    -   (b) adding one or more biocompatible polymers to (a);    -   (c) adding at least one cross-linking agent to (b);    -   (d) cooling (c) and lyophilizing.

In aspects of the invention, the protein of (a) is gelatin and in otheraspects is a mixture of gelatin and collagen. A plasticizer mayadditionally be added to (b) and/or (c). Furthermore, one or more of apharmaceutical, chemical or other agent may be added before and/or afterthe cross-linking reaction of step (c). For example, silver ions,metallic silver or a silver salt may be added to the matrix for thetreatment of burns or other trauma to minimize and/or inhibit infection.The matrix may be fabricated as a covering such as wound barrier, wounddressing, vascular wrap and combinations thereof. The matrix may also befabricated as a sponge. The matrix of the invention may be further usedto reduce chronic inflammation, absorb exudates and/or promote a moistwound environment. The matrix of the invention may also haveincorporated therein a chemical or other agent that is delivered to adesired tissue for at least one of exfoliation and treatment of agerelated conditions in mammals.

In aspects of the invention, the non-adhesive elastic gelatin matrix maybe formed around a web or fibril support. Still in other aspects of theinvention, the gelatin matrix of the invention may be used inconjunction with polymer sheets, films, threads or membranes and meshesof silicone, polyurethane, polyethylene, Dacron™, nylon, silk, celluloseand mixtures thereof.

In other aspects of the present invention, the non-adhesive elasticgelatin matrix may be provided as an occlusive device comprising anocclusive structure and the substantially non-adhesive gelatin matrix,wherein the matrix has opposing surfaces such that one surface of thematrix is affixed to one surface of the occlusive structure with theother surface of the matrix being adapted to cover and be in contactwith tissue. In such aspects the occlusive device may be a plastic filmfor example.

Other features and advantages of the present invention will becomeapparent from the following detailed description. It should beunderstood, however, that the detailed description and the specificexamples while indicating embodiments of the invention are given by wayof illustration only, since various changes and modifications within thespirit and scope of the invention will become apparent to those ski liedin the art from the detailed description.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is an HPLC/UV standard curve of sirolimus.

FIG. 2 shows sirolimus release from a gelatin wrap. S3-1, S3-2 and S3-3were triplicate experiments of samples and every point was the averageof duplicate HPLC analysis.

FIG. 3 shows average drug release from a gelatin wrap.

DETAILED DESCRIPTION OF THE EMBODIMENTS

The present invention is a novel substantially non-adhesive elasticprotein containing matrix that is biocompatible. In embodiments of theinvention, the matrix is an elastic gelatin matrix that retains itselasticity such that it can be configured in a variety of manners as isrequired in various clinical conditions. As such the elastic gelatinmatrix of the invention is flexible and has a variety of clinical usesfor the treatment of a variety of conditions. Furthermore, the matrix ofthe invention can be configured into a variety of formats and used inconjunction with other devices or structures. The matrix of theinvention may also have a variety of pharmaceuticals, chemicals and/orother agents incorporated into the matrix as desired for a variety ofclinical applications. The gelatin matrix of the invention is alsosubstantially non-adhesive meaning that the matrix does notsubstantially adhere to wounds, tissues (such as but not limited toskin, blood vessels and bone) and organs. Lastly, the matrix of theinvention is absorbent, stable in vivo and in vitra and can befabricated with a tensile strength and elasticity as required for adesired clinical application.

The elastic matrix of the invention is made from a solution comprisingprotein or a mixture of proteins, biocompatible polymer(s),cross-linking agent(s), solvent(s) and optionally plasticizer(s). Theprotein for use in the invention is selected from collagen, denaturedcollagen (i.e. gelatin) and mixtures thereof that are further admixedtogether with one or more biocompatible polymers. Plasticizers may beoptionally added. The solution is lyophilized to essentially remove anysolvent and provide the non-adhesive elastic gelatin matrix of theinvention.

In aspects of the invention the biocompatible polymers for use in theinvention are selected from the group consisting of polyethylene glycol(PEG), poly-L-lysine, poly-D-lysine, alginate, chitosan, hyaluronicacid, chondroitin sulfate and mixtures thereof.

In aspects of the invention the cross-linking agents for use in theinvention are selected from the group consisting of1-[3-(dimethylamino)propyl]-3-ethyl carbodiimide (EDC),N-hydroxysuccinimide (NHS), formaldehyde, glutaraldehyde,polyaziridines, diglycidyl ethers and mixtures thereof. In embodiments,EDC and NHS are used in combination as the cross-linking agent.

In aspects of the invention the optional plasticizer for use in thepresent invention is selected from the group consisting of glycerol,propylene glycol, sorbitol and mixtures thereof.

In aspects of the invention the solvent for use in the present inventionis selected from the group consisting of water, methanol, ethanol,isopropanol, dimethylsulfide (DMS), and mixtures thereof. In aspects ofthe invention the solvent is water or if used in addition with a polarorganic solvent, the solvent ratio is typically about 9:1, water topolar organic solvent, in the final mixture.

The solution of collagen and/or gelatin together with the biocompatiblepolymer(s), cross-linking agent(s), optional plasticizer(s) andsolvent(s) are mixed and frozen, and then freeze-dried (i.e.lyophilized). The matrices so produced are substantially non-adhesiveand elastic.

The ranges for the amounts of protein (collagen or gelatin), the amountof biocompatible polymer, the amount of solvent and the amount ofoptional plasticizer in the composition solution and in the lyophilizedfinal composition may be as follows:

% by weight in lyophilized % by weight in initial non-adhesive elasticgelatin composition matrix, lyophilized Protein 0.5-30%  50-90%-collagen or denatured collagen (i.e. gelatin) Biocompatible polymer upto about 10% up to about 40% (in aspects about 10-40%) Cross-linkingagent 0.1-2% 0.5-5% Solvent  50-99% up to about 10% Plasticizer(optional)   0-5%   0-10% *Note that the ranges provided in the tableabove for each of the protein, biocompatible polymer, cross-linkingagent, solvent and plasticizer includes any sub ranges of the rangeslisted.

The non-adhesive elastic gelatin matrix of the invention is made bycombining solutions of protein, biocompatible polymers and cross-linkingagents in a suitable solvent. The solutions of the various constituentsof the matrix as described in the examples are aqueous, however, inaspects of the invention the biocompatible polymer(s) and cross-linkingagent(s) may be first dissolved in a polar organic solvent such as butnot limited to methanol, ethanol, isopropanol and dimethyl sulfoxide(DMSO), and in such a case, the solvent ratio of the final mixture istypically 9:1 in favor of water.

In aspects of the invention, solutions of the protein and biocompatiblepolymer are mixed and incubated for a time period at a temperature suchthat the protein does not set, i.e. form a gel. Temperatures of about55° C. are suitable for such incubation. Should polyethylene glycol(PEG) be used as a biocompatible polymer, the PEG is typically added tothe mixing and incubating protein solutions. A solution of cross-linkingagent is then added and the resultant mixture is mixed for a period oftime until a consistent gel is formed. The gel is then poured into asuitable mold and kept at room temperature for about 10 minutes andsubsequently cooled at about 4° C. for a time period ranging from about30 minutes up to about 12 hours. The mixture is washed to remove solublecomponents such as residual PEG and cross-linking agent and lyophilizedfor a period of about 24 hours or more until the final moisture contentof the non-adhesive elastic gelatin matrix is less than about 10%. It isunderstood by one of skill in the art that the final moisture content ofthe resultant matrix (lyophilized matrix) can be adjusted to any valueup to about 10% by weight of the matrix.

The non-adhesive elastic gelatin matrix of the invention can bedeveloped to be drug-eluting on tissues and organs, including but notlimited to, skin, tissue lacerations, surgical wounds of skin, tissueand organs, blood vessels and bone. In this embodiment, the matrix canbe fabricated to have pharmaceutical, chemical and/or other agentincorporated therein. In one aspect, the pharmaceutical, chemical and/orother agent can be incorporated in an amount of about 0.1% to about 10%into the initial formulation step or about 1.0% to about 25% afterdrying. In another aspect the lyophilized matrix can be soaked in asolution of the desired pharmaceutical, chemical and/or other agent andthen the matrix may be re-lyophilized. Desirable agents for use inconjunction with the present invention include but are not limited tosilver ions, metallic silver or a silver salt, chlorohexidine,triclosan, povidone-iodine, other antimicrobial metals such as copper,platinum, gold, bismuth-based compounds, anesthetics such as lidocaine,antibiotics, immunosuppressants, antiproliferative agents,anti-inflammatory agents, antivirals and combinations thereof. Theselection of the agent for use with the matrix of the invention willdepend on its end use. For example, if used for the treatment ofage-related conditions, an anti-ageing agent may be provided to thematrix before and/or after lyophilization. Such an anti-aging agent maybe selected to treat for example fine lines, wrinkles and skindiscolorations. In this aspect a suitable anti-aging agent may beselected from exfoliants, Vitamin A, Vitamin C, etc. If used to treatbrown spots or discoloration an agent such as hydroquinone may besuitable for incorporation into the matrix.

The non-adhesive elastic gelatin matrix of the invention can befabricated in a variety of forms such as a film, sheet, tube or sponge.The matrix of the invention may be used for example but not limited to awound barrier, wound dressing, vascular wrap and combinations thereof.The matrix of the invention may also be used to reduce chronicinflammation, absorb exudates and/or promote a moist wound environment.The matrix of the invention can also be fabricated as a device inconjunction with other materials such as but not limited to polymersheets, films, threads, membranes or meshes of silicone, polyurethane,polyethylene, Dacron™, nylon, silk, cellulose and combinations thereof.This may be carried out by chemically modifying the surface of the othermaterial by methods of gamma irradiation, plasma or corona dischargeand/or by UV light so that reactive groups are introduced onto thesurface of the material. The reactive groups can then be covalentlylinked to complementary reactive groups present on the non-adhesiveelastic matrix of the present invention by methods as described in theApplicants International Application No. PCT CA02/00246 (the disclosureof which is incorporated herein by reference in its entirety).

The above disclosure generally describes the present invention. A morecomplete understanding can be obtained by reference to the followingspecific Examples. These Examples are described solely for purposes ofillustration and are not intended to limit the scope of the invention.Changes in form and substitution of equivalents are contemplated ascircumstances may suggest or render expedient. Although specific termshave been employed herein, such terms are intended in a descriptivesense and not for purposes of limitation.

EXAMPLES

Without intending to be limiting in scope, the following examples serveto illustrate various embodiments of the invention.

Materials

Gelatin was purchased from Vyse Gelatin Company (5010 North Rose St.,Schiller Park, Ill. 60176). Gelatin (300 Bloom) was obtained by thepartial hydrolysis of a collagen derived from porcine sources: skin,white connective tissues and bones of animals and processed to yield apharmaceutical grade gelatin. Sodium alginate (sodium salt) fromMacrocystis pyrifera (high viscosity, 2% solution at 25° C. approx14,000 cps) was purchased from Sigma Co. (St. Louis, Mo.).Polyethyleneglycol (Average Mn=3,400), 1-[3-(dimethylamino)propyl]-3-ethylcarbodiimide hydrochloride (EDC) andN-hydroxysuccinimide (NHS) and silver lactate were purchased fromAldrich Co. (Milwaukee, Wis. 3201).

Example 1 (Gelatin+Alginate+PEG 3400) Dressing

Stock Solutions:

Gelatin: 20% (w/v)

Alginate: 3.12% (w/v)

PEG 3400: 63% (w/v)

EDC/NHS: 40% (w/v) and 6% (w/v), respectively, (molar ratio: 4/1)

Composition of Water-Saturated Matrix:

Gelatin: 10% (w/w)

Alginate: 1.1% (w/w)

PEG: 7.2% (w/w)

Matrix Synthesis (e.g. to Form About 200 ml of Gel):

Gelatin (100 mL) and alginate (71.2 mL) solutions were mixed andincubated at 55° C. for 30 min. Then 30 mL of PEG 3400 solution wasadded and mixed. Afterward, 5 mL of fresh EDC/NHS solution was added tothe mixture during mixing for 30 seconds. The about 200 ml of gel waspoured into a mold and kept at room temperature for 10 min andsubsequently cooled at 4° C. for 30 min. The matrix was washed in waterovernight to remove soluble components such as PEG and cross-linkingagent. Finally, the matrix was lyophilized. Typically, 0.32 ml of gelmixture yields 1 cm² of matrix surface area.

Example 2 (Gelatin+Alginate+PEG 3400+Glycerol Dressing

Stock Solution:

Gelatin: 20% (w/v)

Alginate: 3.12% (w/v)

PEG 3400: 63% (w/v)

EDC/NHS: 40% (w/v) and 6% (w/v), respectively, (molar ratio: 4/1)

Glycerol: 10% (w/v)

Composition of Water-Saturated Matrix:

Gelatin: 10% (w/w)

Alginate: 1.1% (w/w)

PEG: 7.2% (w/w)

Poly-Lysine: 0.1% (w/w)

Matrix Synthesis (e.g. 200 ml of Gel)

Gelatin (100 mL) and alginate (71.2 mL) solutions were mixed andincubated at 55° C. for 30 min. Then 30 mL of PEG 3400 solution wasadded and mixed. Afterward, 5 mL of fresh EDC/NHS solution was added tothe mixture during mixing for 30 seconds. The about 200 ml of gel waspoured into a mold and kept at room temperature for 10 min andsubsequently cooled at 4° C. for 30 min. The matrix was washed in waterovernight to remove soluble components such as PEG and cross-linkingagent. Usually, 0.32 ml of gel mixture yields 1 cm² of matrix surfacearea. The matrix was soaked in 2.85 L of 10% glycerol (4.4 ml ofglycerol per 1 cm² of gel). Finally the matrix was lyophilized.

Example 3 (Gelatin+Alginate+PEG 3400+Poly-L-lysine) Dressing

Stock Solution:

Gelatin: 20% (w/v)

Alginate: 3.12% (w/v)

PEG 3400: 63% (w/v)

Poly-L-Lysine: 2% (w/v)

EDC/NHS: 40% (w/v) and 6% (w/v), respectively, (molar ratio: 4/1)

Composition of Water-Saturated Matrix:

Gelatin: 10% (w/w)

Alginate: 1.1% (w/w)

PEG: 7.2% (w/w)

Poly-lysine: 0.1% (w/w)

Matrix Synthesis

Gelatin and alginate solutions were mixed as in previous examples. Then10 ml of 2% poly-L-lysine was added with mixing. 30 ml of PEG 3400 wasadded to the mixture and mixed. The remaining procedures to yield thefinal matrix were identical to previous examples.

Example 4 (Gelatin+Alginate+PEG3400+Poly-L-lysine+Glycerol Dressing

Stock Solution:

Gelatin: 20% (w/v)

Alginate: 3.12% (w/v)

PEG 3400: 63% (w/v)

Poly-L-Lysine: 2% (w/v)

EDC/NHS: 40% (w/v) and 6% (w/v), respectively, (molar ratio: 4/1)

Glycerol: 10% (w/v)

Composition of Water-Saturated Matrix:

Gelatin: 10% (w/w)

Alginate: 1.1% (w/w)

PEG: 7.2% (w/w)

Poly-L-lysine: 0.1% (w/w)

Matrix Synthesis

Gelatin and alginate solutions were mixed as in previous examples. Then10 ml of 2% poly-L-lysine was added with mixing. 30 ml of PEG 3400 wasadded to the mixture and mixed. The remaining procedures to yield thehydrated and washed matrix were identical to previous examples. Finally,the matrix was soaked in 2.85 L of 10% glycerol (4.4 ml of glycerol per1 cm² of gel) and then lyophilized.

Example 5 (Gelatin+Alginate) Dressing

Stock Solution:

Gelatin: 1% (w/v)

Alginate: 1% (w/v)

EDC/NHS: 40% (w/v) and 6% (w/v), respectively, (molar ratio: 4/1)

Composition of Water-Saturated Matrix:

Gelatin: 0.9% (w/w)

Alginate: 0.1% (w/w)

Matrix Synthesis (e.g. 200 ml of Gel)

Gelatin (180 mL) and alginate (20 mL) solutions were mixed and incubatedat 55° C. for 30 min. Afterward 1.4 ml of fresh EDC/NHS solution wasadded to the mixture with mixing for 1 minute. The mixture was pouredinto a mold (10 ml per 20 cm² area), and then frozen at −20° C.overnight. Finally, the matrix was lyophilized.

The various matrices as in Examples 1-5 can be poured (prior to theinitial cooling/freezing step) directly over other materials (withoutmodification as outlined above), such as but not limited to polymersheets, films, threads, membranes or meshes of silicone, polyurethane,polyethylene, Dacron™, nylon, silk, cellulose and combinations thereofthat may or may not contain added agents, such as therapeutics,silver-containing materials or other medicaments so as to embed themesh, net or fibers within the matrices of examples 1-5.

Example 6 In Vitro Degradation of Gelatin Sponge by Collagenase

The degradation of cross-linked gelatin-based materials was studiedusing bacterial collagenase. The collagenase used in this study was fromClostridium (EC 3.4.24.3) and had an activity of 362 U/mg solid. Samples(50 mg) were incubated at 37° C. in 10 mL of collagenase solution withconcentration of 15.35 U/mg in 0.1M Tris-HCl buffer (pH=7.4) containing0.005M CaCl₂ and 0.05 mg/mL sodiumazide. After each incubation interval,the sample was carefully washed three times with de-ionized water andlyophilized overnight. The extent of degradation was expressed as thepercentage of weight remaining after degradation.

The biodegradation of gelatin-based dressings in accordance with thepresent invention was rapid in the presence of collagenase, as would beexpected. For nearly all of the matrices, approximately 80% of thegelatin sponge was degraded in the first 24 hours. The gelatin matrixcontaining gelatin and alginate appeared less affected by the action ofcollagenase (Table 1).

TABLE 1 Biodegradation of gelatin dressings at specified times(expressed as % of weight remaining) in the presence of collagenaseSample Matrix 0 hr 24 hr 48 hr 72 hr 1% Gelatin + Alginate 100 96.1 75.965.4 10% Gelatin + PEG + Alginate 100 21.0 22.2 17.4 10% Gelatin + PEG +Alginate + Glycerol 100 23.3 12.8 21.8 10% Gelatin + PEG + Alginate +Glycerol + 100 18.4 14.3 14.4 Poly-L-Lysine 10% Gelatin + PEG +Alginate + Poly-L-Lysine 100 16.5 14.8 12.6

Example 7 Determination of Water Uptake Ability

To determine the water uptake ability of the dressing, about 40 mg ofdressing was placed in a 20 ml glass vial to which was added 15 ml ofwater or PBS (0.01M, pH=7.4). The sample was weighed after hydration for5 min at room temperature and then incubated at 37° C. for 4 hr and 24hr. The sample was weighed at both incubation intervals. The wateruptake ability is expressed as the weight ratio of absorbed water to drydressing (Table 2). Matrices particularly without plasticizer are allvery absorbent and can soak up to approximately 30-35 times their weightin water in about 24 hours.

TABLE 2 Water uptake at specified times, grams of water/grams of matrix(g/g) 5 min 4 min 24 min Sample Matrix (g/g) (g/g) (g/g) 1% Gelatin +Alginate 131.6 — 28.7 10% Gelatin + PEG + Alginate 16.2 27.3 32.1 10%Gelatin + PEG + Alginate + Glycerol 2.5 10.8 11.7 10% Gelatin + PEG +Alginate + Poly-L-Lysine 9.0 29.9 34.6 10% Gelatin + PEG + Alginate +Poly-L-Lysine + 1.8 9.6 10.5 Glycerol * values are a ratio, value: 1.

Example 8 Determination of Water Solubility

To determine the water solubility of the dressing, place a sample with aknown weight was placed into a glass vial to which 15 mls of water wasadded. This was incubated at 40° C. for 24 hr and then the water removedand the sample washed twice with water. The sample was then dried in a100° C. oven to a constant weight and then re-weighed. The watersolubility is typically expressed as percentage of weight lost afterwater treatment (Table 3). The matrices exhibited a good watersolubility providing an indication of its clinical longevity. Lowersolubility resulted in a more stable matrix at body temperature. Thussuch matrices can be removed from a wound for example as a single sheetafter 24 hours or more with minimum patient discomfort. While allmatrices demonstrated good stability, those without plasticizer wereeven more stable.

TABLE 3 Stability of 10% gelatin-based materials after incubation in 40°C. water for 24 hr. Weight Water W1 W2 Remaining Solubility SampleMatrix (g) (g) (g) (%) 1% Gelatin + Alginate 0.0315 0.0260 83.4 16.6 10%Gelatin + PEG + Alginate 0.0335 0.0261 77.9 22.1 10% Gelatin + PEG +Alginate 0.0308 0.0235 76.3 23.7 10% Gelatin + PEG + Alginate + Glycerol0.0448 0.0107 23.9 76.1 10% Gelatin + PEG + Alginate + Glycerol 0.04430.0108 24.4 75.6 10% Gelatin + PEG + Alginate + Poly-L-Lysine 0.03640.0305 83.8 16.2 10% Gelatin + PEG + Alginate + Poly-L-Lysine 0.03610.0294 81.4 18.6 10% Gelatin + PEG + Alginate + Poly-L-Lysine + 0.05040.0115 22.82 77.2 Glycerol 10% Gelatin + PEG + Alginate +Poly-L-Lysine + 0.0507 0.0124 24.5 75.5 Glycerol Note: W1: weight oforiginal sample W2: weight of dried sample after treatment WeightRemaining = (W2/W) × 100% Water Solubility = (1 − W2/W1) × 100%

Example 9 Determination of Mechanical Properties

The mechanical properties were determined for the tensile strength andelongation at break of hydrated gelatin matrices (4.5 cm×1 cm), whichhad been soaked in 0.01M, pH=7.4 PBS buffer solution for 1 hour at roomtemperature. A 0.5 kg load cell was used with an extension rate of 5mm/min (Table 4). This example demonstrates that tensile strength andelasticity of the matrices can be adjusted through the variation of theamount and combination of components of the matrix. This is valuable forproviding matrices for a variety of clinical applications such as anartificial skin or wound dressing.

TABLE 4 Mechanical properties of gelatin-based dressings TensileStrength Elongation at Sample No. (g/cm²) Break (%) 1% Gelatin +Alginate 25.5 53.0 10% Gelatin + PEG 30.5 126.8 10% Gelatin + PEG +Alginate 73.8 174.9 10% Gelatin + PEG + Alginate + 83.8 155.4 Glycerol10% Gelatin + PEG + Alginate + 111.4 176.3 Poly-L-Lysine 10% Gelatin +PEG + Alginate + 148.8 193.3 Poly-L-Lysine + Glycerol

Example 10 Addition of Sirolimus to Gelatin-Based Dressings

Examples 1-5 are directed to the manufacture of a range materials thatmay be used as carriers for various medicaments such as theantiproliferative and anti-inflammatory drug Sirolimus (Rapamycin) fordelivery to the surface of tissues and organs. While the drug Sirolimusis used in this example, it is noted that other drugs may be used eitheralone or in combination to be delivered to the surface of tissues andorgans.

Stock Solution:

Gelatin: 1% to water

Alginate: 1% to 0.05N NaOH

EDC/NHS: 400 mg/60 mg per 1 ml of 10 mM of Mes saline solution (pH=4.5)

Glycerol: 1% (in water).

The Formulation in the Matrix:

Gelatin: 0.9% (w/v)

Alginate: 0.1% (w/v)

Water

Procedure:

Preparation of Stock Solutions

Solution 1: 2 g of gelatin was added to 180 ml of water; after gelatinwas completely hydrated in water, the mixture was incubated at 50° C. todissolve the gelatin.

Solution 2: 0.2 g of alginate was added into 20 ml of 0.05N NaOH, andincubated at 50° C. to dissolve the alginate.

Solution 3: 400 mg of EDC+60 mg of NHS was added into 1 ml of 10 mM Messaline solution (pH=4.5).

Solution 4: 2 g of glycerol was added into 200 ml of water

Matrix Synthesis (e.g. 200 ml of Gel)

-   -   1. Solution 1 and Solution 2 were mixed and incubated at 55° C.        for 30 min. Afterward, 1.379 ml of fresh EDC/NHS solution was        added to mixture with mixing for 1 minute. The mixture was        poured into a mold (9 ml per 20 cm² a, then froze at −20 degrees        ° C. for overnight.    -   2. Cross-linked gel was washed by Milli-Q water 4 times with 500        ml per matrix (Refresh water every 1 hr); the matrix was        refrozen at room temperature, soaked in 9 ml of 1% glycerol per        20 cm² gel. Then gel was frozen again at −20° C. and        lyophilized. The matrix may be prepared as a thin film.        Sirolimus Loading of Matrices:

The above matrix, (or any of the matrices of examples 1-5) was cut into1×4 cm square. Afterwards, 80 ul of drug solution (1 mg/80 ul ethanol)was added on the surface of the material, or the material was immersedin a solution of drug. After drug loading into the film, the film wasallowed to dry and the drug-loaded film was exposed to gamma-irradiationfor 8 hr (25 KGy).

In-Vitro Drug Release Study

Procedure:

1. Preparation of Samples

-   -   1) One matrix (1×4 cm) was cut into equal 3 pieces.    -   2) Dissolve 1 mg of drug in 90 ul of ethanol.    -   3) Adding 30 ul of drug solution into each matrix piece of step        1.    -   4) Evaporate Ethanol.        2. Drug Release Study    -   1) Place samples in 15 ml of Falcon tubes.    -   2) Add 5 ml of PBS (10 mM, pH=7) to each tube.    -   3) Incubate tubes at 37° C. for 10 days.    -   4) Withdraw samples of 2.5 ml at intervals of 3 hr, 1 day, 3        days, 5 days, 7 day, 9 days, and 2.5 ml fresh PBS was replaced        for samples taken.

5) HPLC analysis of samples.

Column: Hypersil ODS, 100×2.1 mm

Flow rate: 0.2 ml/min

Detection: UV, 278 nm

Mobile phase: 600 ml acetonitrile and 400 ml of water

Temperature: 50° C.

Injection volume: 10 μl

Sirolimus Standards: 0 ug/ml, 1 ug/ml, 2.5 ug/ml, 5 ug/ml, 10 ug/ml

Analysis: Each sample was injected two times for analysis.

Although preferred embodiments of the invention have been describedherein in detail, it will be understood by those skilled in the art thatvariations may be made thereto without departing from the spirit of theinvention or the scope of the appended claims.

1. A non-adhesive elastic protein matrix comprising: (a) about 50-90% byweight of one or more proteins selected from the group consisting ofcollagen, denatured collagen and mixtures thereof; (b) about 10-40% byweight of one or more biocompatible polymers selected from the groupconsisting of polyethylene glycol, poly-L-lysine, alginate, chitosan,hyaluronic acid, chondroitin sulfate and mixtures thereof; (c) about0.5-5% by weight of one or more cross-linking agents; and (d) one ormore pharmaceutical, chemical and other agents; wherein said matrix islyophilized after addition of (c) to result in a matrix that isnon-adherent to wounds, tissues and organs and can be removed after 24hours.
 2. The matrix of claim 1, wherein said matrix additionallycomprises one or more plasticizers.
 3. The matrix of claim 1, whereinsaid cross-linking agent is selected from the group consisting of1-[3-(dimethylamino)propyl]-3-ethyl carbodiimide (EDC),N-hydroxysuccinimide (NHS), formaldehyde, glutaraldehyde,polyaziridines, diglycidyl ethers and mixtures thereof.
 4. The matrix ofclaim 3, wherein said cross-linking agent is a mixture of1-[3-(dimethylamino)propyl]-3-ethyl carbodiimide (EDC) andN-hydroxysuccinimide (NHS).
 5. The matrix of claim 1, wherein saidmatrix contains less than about 10% moisture content.
 6. The matrix ofclaim 2, wherein said plasticizer is provided in an amount of up toabout 10% by weight of said matrix.
 7. The matrix of claim 1, whereinsaid one or more pharmaceutical, chemical and other agent is selectedfrom the group consisting of antibiotic, antiviral, chlorhexidine,triclosan, povidone-iodine, antimicrobial metals, bismuth-basedcompounds, immunosuppressants, anti-proliferative agents,anti-inflammatory agents, anesthetics, anti-aging agents and mixturesthereof.
 8. The matrix of claim 7, wherein said antimicrobial metal isselected from the group consisting of silver ions, metallic silver,silver salt, copper, platinum, gold and mixtures thereof.
 9. The matrixof claim 7, wherein said anesthetic is lidocaine.
 10. The matrix ofclaim 1, wherein said one or more pharmaceutical, chemical and otheragent is provided in an amount of about 1% to about 25% by weight ofsaid matrix.
 11. The matrix of claim 1, wherein said matrix is providedas a film, sheet, tube or sponge.
 12. The matrix of claim 1, whereinsaid matrix is provided together with a material selected from the groupconsisting of polymer sheets, polymer films, threads, siliconemembranes, silicone meshes, polyurethane, polyethylene, polyethyleneterephthalate, nylon, silk, cellulose and combinations thereof.
 13. Thematrix of claim 1, wherein said matrix is provided as a wound dressing,wound barrier, tissue cover and/or vascular cover.
 14. The matrix ofclaim 1, wherein said matrix reduces chronic inflammation.
 15. Thematrix of claim 1, wherein said matrix absorbs exudates.
 16. The matrixof claim 1, wherein said matrix promotes a moist environment.
 17. Thematrix of claim 1, wherein said pharmaceutical agent is sirolimus.